Vibrio harveyi

  • 文章类型: Journal Article
    象牙壳(Babyloniaareolata)是最有前途的高品质海产品之一。然而,象牙壳在培养期间容易感染哈维氏弧菌。在这项研究中,我们调查了生化指标,PBS对照组(PC)和感染组(A3)在24小时后1×109CFU/mLV.harveyi的象牙壳肝胰腺的组织学变化和转录组反应。结果表明,与PC组相比,生化指标,包括丙二醛(MDA),活性氧(ROS),酸性磷酸酶(ACP),和Caspase3(Casp-3)在A3组感染弧菌24h后显著升高(p<0.05)。A3组肝胰腺损伤严重,组织的柱状上皮细胞增大,消化细胞的空间增加了,出现了空泡腔。在感染组中总共获得了95,581个单基因,并鉴定了2,949个(1,787个上调和1,162个下调)差异表达基因(DEGs)。GO和KEGG富集分析表明,DEGs主要富集在免疫系统过程中(GO:0002376),抗氧化活性(GO:0016209),溶酶体(ko04142),Toll和IMD信号通路(ko04624),和其他人。这些生物学功能和途径与免疫和炎症反应以及细胞凋亡有关。随机选择12个DEGs进行实时定量PCR(RT-qPCR)验证,这些DEGs的表达谱与转录组数据一致,确认转录组结果的准确性和可靠性。总之,V.哈维伊象牙壳感染诱导氧化应激,导致严重的肝胰腺损伤,刺激谷胱甘肽的产生以中和过量的ROS,和抗菌肽的生产,以抵消细菌感染的有害影响,反过来改变免疫和炎症反应,最终导致细胞凋亡。本研究为探索哈维氏弧菌感染后的免疫调节机制提供了有价值的信息,并为预防哈维氏弧菌感染提供了分子依据。
    The ivory shell (Babylonia areolata) is one of the most promising high quality marine products. However, ivory shell is susceptible to Vibrio harveyi infection during the culture period. In this study, we investigated the biochemical indicators, histological changes and transcriptomic response in the hepatopancreas of ivory shells from the PBS control group (PC) and infection group (A3) with 1 × 109 CFU/mL V. harveyi after 24 h. Results showed that compared to the PC group, biochemical indicators, including malondialdehyde (MDA), reactive oxygen species (ROS), acid phosphatase (ACP), and Caspase 3 (Casp-3) were significantly increased (p < 0.05) in A3 group after V. harveyi infection for 24 h. Compared with the PC group, the hepatopancreas of A3 group were seriously damaged, the columnar epithelial cells of the tissue were enlarged, the space of digestive cells was increased, and vacuolar cavities appeared. A total of 95,581 unigenes were obtained and 2949 (1787 up-regulated and 1162 down-regulated) differential expressed genes (DEGs) were identified in the infecting group. GO and KEGG enrichment analysis showed that DEGs were mainly enriched in immune system process (GO:0002376), antioxidant activity (GO:0016209), lysosome (ko04142), Toll and IMD signaling pathway (ko04624), and others. These biological functions and pathways are associated with immune and inflammatory responses and apoptosis. Twelve DEGs were randomly selected for real-time quantitative PCR (RT-qPCR) validation, and the expression profiles of these DEGs were consistent with the transcriptome data, confirming the accuracy and reliability of the transcriptome results. In summary, V. harveyi infection of ivory shells inducing oxidative stress, leading to severe hepatopancreatic damage, stimulating glutathione production to neutralize excessive ROS, and antimicrobial peptides production to counteract the deleterious effects of bacterial infection, which in turn modifying the immune and inflammatory response, ultimately resulting in apoptosis. This study provided valuable information to explore the immune regulation mechanism after V. harveyi infection and provides molecular basis to support the prevention of V. harveyi infection.
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  • 文章类型: Journal Article
    哈氏弧菌是天然海洋生境中的正常菌群,是海洋动物中的重要机会病原体。这种细菌感染海洋动物后可引起一系列病变,其中肌肉坏死和溃疡是最常见的症状。本研究探索了哈维伊氏弧菌从海水环境到寄主鱼类肌肉环境的适应机制。综合转录组分析显示,在适应宿主鱼类肌肉环境的过程中,哈维伊氏弧菌的转录组发生了巨大变化。基于基因本体论(GO)和京都基因和基因组百科全书(KEGG)分析,鞭毛组装,氧化磷酸化,细菌趋化性,两组分系统在哈维伊氏弧菌对宿主鱼肌肉的适应中起着至关重要的作用。生物表型的比较表明,哈维伊弧菌显示鞭毛长度显着增加,游泳,抽搐,趋化性,附着力,和宿主鱼肌肉诱导后的生物膜形成,和它的主要氨基酸,特别是宿主肌肉诱导的细菌趋化,Ala和Arg.可以推测,氨基酸诱导的细菌趋化性增强在哈氏弧菌从海水到宿主鱼肌肉的适应中起着关键作用。
    Vibrio harveyi is a normal flora in natural marine habitats and a significant opportunistic pathogen in marine animals. This bacterium can cause a series of lesions after infecting marine animals, in which muscle necrosis and ulcers are the most common symptoms. This study explored the adaptation mechanisms of V. harveyi from the seawater environment to host fish muscle environment. The comprehensive transcriptome analysis revealed dramatic changes in the transcriptome of V. harveyi during its adaptation to the host fish muscle environment. Based on Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) analysis, flagellar assembly, oxidative phosphorylation, bacterial chemotaxis, and two-component systems play crucial roles in V. harveyi\'s adaptation to host fish muscle. A comparison of biological phenotypes revealed that V. harveyi displayed a significant increase in flagellar length, swimming, twitching, chemotaxis, adhesion, and biofilm formation after induction by host fish muscle, and its dominant amino acids, especially bacterial chemotaxis induced by host muscle, Ala and Arg. It could be speculated that the enhancement of bacterial chemotaxis induced by amino acids plays a key role in the adaptation of V. harveyi from seawater to the muscle of the host fish.
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  • 文章类型: Journal Article
    Toll样受体(TLR)代表了模式识别受体的一个重要类别,已对其在抵抗病原体入侵中的关键作用进行了广泛研究。尽管如此,在C.altivelis中,明显缺乏与TLR家族基因相关的免疫反应的全面鉴定和探索。本研究成功鉴定并命名了14个基因:Catlr1-1,Catlr1-2,Catlr2-1,Catlr2-2,Catlr3,Catlr5,Catlr7,Catlr8,Catlr9,Catlr13-1,Catlr13-2,Catlr18,Catlr21和Catlr22。进行了一系列生物信息学分析,包括蛋白质特性的分析,检查基因结构,进化评估,和蛋白质三级结构的预测。分析了Catlr基因在五种免疫组织中的表达模式:肝脏,脾,脾肾,吉尔,和肠,在健康和细菌刺激的鱼类中。结果表明,不同组织和不同基因在哈氏弧菌感染后表现出不同的表达模式,表明所有Catlr成员参与各种组织感染后的免疫反应。此外,免疫组织的组织学评估揭示了不同程度的损伤。总之,这项对TLR基因家族的研究提供了新的信息,有助于更深入地理解C.altivelis的免疫应答机制。
    Toll-like receptors (TLRs) represent a prominent category of pattern recognition receptors that have been extensively investigated for their pivotal role in combating pathogen incursions. Despite this, there has been a notable absence of comprehensive identification and exploration of the immune response associated with the TLR family genes in C. altivelis. This study successfully identified and named fourteen genes as Catlr1-1, Catlr1-2, Catlr2-1, Catlr2-2, Catlr3, Catlr5, Catlr7, Catlr8, Catlr9, Catlr13-1, Catlr13-2, Catlr18, Catlr21, and Catlr22. A series of bioinformatic analysis were performed, encompassing analysis of protein properties, examination of gene structures, evolutionary assessments, and prediction of protein tertiary structures. The expression patterns of Catlr genes were analyzed in five immune tissues: liver, spleen, kidney, gill, and intestine, in both healthy and bacterial stimulated-fish. The results showed that different tissue and different genes showed differed expression patterns after V. harveyi infection, indicating the involvement of all Catlr members in mounting immune responses following infection in various tissues. Additionally, histological evaluations of immune tissues unveiled varying levels of damage. In conclusion, this investigation into the TLR gene family offers novel information that contribute to a more profound comprehension of the immune response mechanisms in C. altivelis.
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  • 文章类型: Journal Article
    如今,欧洲鲈鱼(Dicentrarchuslabrax)水产养殖正在经历重大扩张。然而,水产养殖业受到弧菌病的困扰。2022年的七个月期间,在法国北部的欧洲海底农场研究了哈维氏弧菌的时空动态。将混凝土试样悬浮,并从不同深度(0.3m,2.15米和4米深),提供对生物膜和浮游V.harveyi动力学的见解。V.harveyi的丰富,在生物膜和自由生活形式中,呈正相关。水参数揭示了温度的季节性波动,pH值,和盐度,在整个水柱中没有观察到显着差异。V.harveyi的定量显示深度之间没有显着差异,但是季节性,在8月份观测到峰值丰度,与温度升高有关。主成分分析将温度确定为主要驱动因素,但也有额外的参数,例如盐度和pH。弧菌病发生在采样期间,提供对以前情况的有价值的见解,during,在爆发之后。这项研究强调了了解V.harveyi行为在水产养殖中的重要性,特别是在全球变暖的背景下,有效的疾病管理和可持续的做法。
    Nowadays, European seabass (Dicentrarchus labrax) aquaculture is undergoing a significant expansion. Nevertheless, the aquaculture industry is plagued by vibriosis. The spatial and temporal dynamics of Vibrio harveyi were studied on a European seabass farm in northern France during seven months of 2022. Concrete specimens were suspended and water was pumped from different depths (0.3 m, 2.15 m and 4 m deep), providing insights into the biofilm and planktonic V. harveyi dynamics. The abundances of V. harveyi, in the biofilm and free-living forms, were positively correlated. The water parameters revealed seasonal fluctuations in temperature, pH, and salinity, with no significant differences observed across the water column. Quantification of V. harveyi revealed no significant differences between depths, but seasonality, with peak abundances observed in August, correlated with temperature increases. Principal component analysis identified temperature as a primary driver, but also additional parameters, such as salinity and pH. Vibriosis occurred during the sampling period, providing valuable insights into the conditions before, during, and after the outbreaks. This study underscores the importance of understanding V. harveyi behaviour in aquaculture, particularly in the context of global warming, for effective disease management and sustainable practices.
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  • 文章类型: Journal Article
    描述了将几丁质酶固定在磁性纳米颗粒(MNP)上,作为生物相容性载体,用于从几丁质废物中酶促生产二-N-乙酰基几丁糖。几丁质酶固定化是可行的,固定化产率为88.9±1.6%,活性保留为97.8±1.0%,与游离酶相比,固定化赋予了更好的热和储存稳定性。作为实际益处,与磁性纳米载体的连接使得在重复施用运行之后能够容易地回收酶,并且因此可持续地重复使用。在固定状态下,几丁质酶在16个反应循环后保留了显着的39.7±2.6%的起始活性。此外,固定化几丁质酶在一步反应中将虾壳和鱿鱼壳几丁质转化为二-N-乙酰基几丁糖方面比游离几丁质酶具有更高的催化活性。来自虾壳的纯化化合物的最终产率为37.0±1.2%,来自鱿鱼几丁质的最终产率为61.1±0.5%。总之,开发了一种有效的基于MNP的几丁质酶固定化系统,具有大规模生产的潜力。
    Described is chitinase immobilization on magnetic nanoparticles (MNPs) as biocompatible support for enzymatic production of di-N-acetyl chitobiose from chitin waste. Chitinase immobilization was feasible with an immobilization yield of 88.9 ± 1.6 % with 97.8 ± 1.0 % retention of activity and compared to free enzyme work, immobilization conferred better thermal and storage stability. As practical benefit the attachment to magnetic nanocarriers enabled easy enzyme recovery after repeated application runs and thus sustainable reuse. In fixed state chitinase retained a remarkable 39.7 ± 2.6 % of the starting activity after 16 reaction cycles. Furthermore, immobilized chitinase showed higher catalytic activity than free chitinase in converting shrimp shells and squid-pens chitins into di-N-acetyl chitobiose in a single-step reaction. The final yield of purified compound was 37.0 ± 1.2 % from shrimp shells and 61.1 ± 0.5 % from squid-pens chitin. In conclusion, an efficient MNP-based chitinase immobilization system with the potential for large-scale production was developed.
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  • 文章类型: Journal Article
    microRNAs(miRNAs)在基因转录后调控中起着重要作用,在调节海洋物种的免疫应答中起着关键作用。特别是在病原体攻击期间。本研究主要研究miR-7562的功能及其对黑虎虾(Penaeusmonodon)感染哈维氏弧菌自噬的调节作用,一种经济上重要的水生物种。我们成功地克隆和鉴定了两个必需的自噬相关基因(ATGs)。PmATG5和PmATG12,然后鉴定可能参与共同调节这些基因的miRNAs,其中值得注意的是miR-7562,miR-8485和miR-278。随后的细菌攻击实验和双荧光素酶报告基因测定确定miR-7562是两个基因的主要调节因子。特别是通过靶向每个基因的3UTR。通过使用模拟物和antagomirs操纵miR-7562的体内水平,我们发现PmATG5和PmATG12的表达存在显着差异,这与自噬活性的改变相对应。值得注意的是,miR-7562过表达导致PmATG5和PmATG12的下调,导致减弱的自噬反应。相反,miR-7562敲低提高了这些基因的表达水平,从而增强自噬活性。我们的研究结果进一步表明,在哈维伊氏弧菌感染期间,miR-7562通过特异性靶向ATG5-ATG12复合物继续影响自噬途径。这项研究不仅揭示了miRNA依赖的调控虾自噬免疫的机制,而且还提出了miR-7562作为旨在增强甲壳类水产养殖业抗病性的治疗策略的有希望的靶标。
    MicroRNAs (miRNAs) play a fundamental role in the post-transcriptional regulation of genes and are pivotal in modulating immune responses in marine species, particularly during pathogen assaults. This study focused on the function of miR-7562 and its regulatory effects on autophagy against Vibrio harveyi infection in the black tiger shrimp (Penaeus monodon), an economically important aquatic species. We successfully cloned and characterized two essential autophagy-related genes (ATGs) from P. monodon, PmATG5 and PmATG12, and then identified the miRNAs potentially involved in co-regulating these genes, which were notably miR-7562, miR-8485, and miR-278. Subsequent bacterial challenge experiments and dual-luciferase reporter assays identified miR-7562 as the principal regulator of both genes, particularly by targeting the 3\'UTR of each gene. By manipulating the in vivo levels of miR-7562 using mimics and antagomirs, we found significant differences in the expression of PmATG5 and PmATG12, which corresponded to alterations in autophagic activity. Notably, miR-7562 overexpression resulted in the downregulation of PmATG5 and PmATG12, leading to a subdued autophagic response. Conversely, miR-7562 knockdown elevated the expression levels of these genes, thereby enhancing autophagic activity. Our findings further revealed that during V. harveyi infection, miR-7562 continued to influence the autophagic pathway by specifically targeting the ATG5-ATG12 complex. This research not only sheds light on the miRNA-dependent mechanisms governing autophagic immunity in shrimp but also proposes miR-7562 as a promising target for therapeutic strategies intended to strengthen disease resistance within the crustacean aquaculture industry.
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  • 文章类型: Journal Article
    哈维氏弧菌导致高死亡率,并严重限制石斑鱼的培养。肠道菌群是抵抗病原体入侵的重要生物屏障。在这项研究中,我们调查了肠道微生物群落的动态变化,在0、3和7天,珍珠龙胆石斑鱼(Epinephelusfuscoguttatusult×Epinepheluslanceolatus)的基因转录和免疫反应特征(称为d0,d3和d7组,分别)感染哈维伊弧菌后。结果表明,d7处理降低了肠道微生物多样性,增加了变形菌和蓝细菌的比例。值得注意的是,几个假定的病原属(鞘氨醇单胞菌和拟杆菌)增殖,而假定的益生菌属(红球菌和乳酸菌)减少,肠道细菌的这些变化可能与宿主免疫相关分子的改变有关。d3和d7治疗也改变了主要与肠道免疫功能相关的组织形态学和基因转录谱,如“MAPK信号通路”,“细胞凋亡”和“Toll样受体(TLR)信号通路”。此外,d3组诱导了抗氧化系统的稳态失调,细胞因子和TLR信号,在d7组中有逐渐恢复正常状态的趋势,伴随着细胞凋亡的过程。致病性感染抑制了JNK通路的表达并增强了ERK通路。总之,由V.harveyi感染期间发生的免疫变化引起的肠道细菌群落的生态失调破坏了珍珠龙胆石斑鱼的肠道健康。这些结果为全面了解鱼类的免疫防御机制,制定石斑鱼养殖的疾病控制策略提供了有价值的参考。
    Vibrio harveyi causes high mortality and severely limits grouper culture. The gut microbiota is an important biological barrier against pathogen invasion. In this study, we investigated dynamic changes in the intestinal microbial community, gene transcription and immune responses signatures of pearl gentian grouper (Epinephelus fuscoguttatus♂ × Epinephelus lanceolatus♀) at 0, 3 and 7 days (referred to as d0, d3 and d7 groups, respectively) after infection with V. harveyi. The results demonstrated that the d7 treatment reduced the gut microbial diversity and increased the proportion of Proteobacteria and Cyanobacteria. Notably, several putative pathogenic genera (Sphingomonas and Bacteroides) proliferated, while putative probiotic genera (Rhodococcus and Lactobacillus) reduced, and these changes in intestinal bacteria might be correlated to the alterations of host immune-related molecules. The d3 and d7 treatments also altered the histomorphology and gene transcription profiles mainly associated with immune function in intestine, such as \'MAPK signaling pathway\', \'Apoptosis\' and \'Toll-like receptor (TLR) signaling pathway\'. Furthermore, d3 group induced a homeostatic dysregulation of the antioxidant system, cytokines and TLR signaling, with a tendency to gradually return to a normal state in d7 group, along with the apoptosis process. The pathogenic infection suppressed the expression of JNK pathway and enhanced the ERK pathway. In conclusion, the dysbiosis of the intestinal bacterial communities caused by the immune changes that occurred during V. harveyi infection disrupted the intestine health in the pearl gentian grouper. These results provided a comprehensive understandings of the immune defense mechanisms in fish and valuable references to develop disease control strategies in grouper aquaculture.
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  • 文章类型: Journal Article
    据报道,哈维氏弧菌分离株K2014767的完整基因组序列是在美国公共展示水族馆的一次特定物种死亡事件中从圈养的加勒比多刺龙虾(Panulirusargus)中分离的。
    The complete genome sequence is reported for Vibrio harveyi isolate K2014767, isolated from a captive Caribbean spiny lobster (Panulirus argus) during a species-specific mortality event in a public display aquarium in the United States.
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  • 文章类型: Journal Article
    沉浸式疫苗接种,尽管比注射免疫更容易管理,有时对抗原摄取有挑战,导致次优保护。在这项研究中,探索了使用富含氧气的纳米气泡的水(ONB)和带正电的壳聚糖(CS)增强亚洲鲈鱼(Latescalcarifer)中热灭活的哈维氏弧菌疫苗抗原摄取的新策略。评估了鱼g中的抗原吸收,与ONB和CS的四种不同疫苗组合的抗体反应和疫苗功效一样,和两个对照组。在引入疫苗之前预先混合ONB和CS,称为(ONB+CS)+VAC,导致优异的抗原摄取和抗V。与其他配方相比,血清和粘液中的harveyi抗体(IgM)产生。口服助推器的积分(4.22×108CFU/g,在第21-25天)的疫苗试验实验中,旨在进一步评估暴露于V.harveyi后的生存率如何提高。在42天内测量抗体反应,通过用V.harveyi进行实验攻击来评估疫苗的效力。免疫相关基因IL1β的表达,TNFα,在攻击后(dpc)1、3和7天评估CD4、CD8、IgT和抗体水平。结果显示,(ONB+CS)+Vac组的抗体水平在浸没免疫和口服加强后始终高于其他组,在用V.harveyi攻击后,免疫相关基因的表达也升高。最终,该组表现出明显更高的相对生存率(RPS),为63%±10.5%,展示了ONB-CS-Vac复合物作为增强抗原摄取的有前途的浸没疫苗接种策略的潜力,刺激免疫反应,提高亚洲鲈鱼对弧菌病的存活率。
    Immersion vaccination, albeit easier to administer than immunization by injection, sometimes has challenges with antigen uptake, resulting in sub-optimal protection. In this research, a new strategy to enhance antigen uptake of a heat-inactivated Vibrio harveyi vaccine in Asian seabass (Lates calcarifer) using oxygen nanobubble-enriched water (ONB) and positively charged chitosan (CS) was explored. Antigen uptake in fish gills was assessed, as was the antibody response and vaccine efficacy of four different combinations of vaccine with ONB and CS, and two control groups. Pre-mixing of ONB and CS before introducing the vaccine, referred to as (ONB + CS) + Vac, resulted in superior antigen uptake and anti-V. harveyi antibody (IgM) production in both serum and mucus compared to other formulas. The integration of an oral booster (4.22 × 108 CFU/g, at day 21-25) within a vaccine trial experiment set out to further evaluate how survival rates post exposure to V. harveyi might be improved. Antibody responses were measured over 42 days, and vaccine efficacy was assessed through an experimental challenge with V. harveyi. The expression of immune-related genes IL1β, TNFα, CD4, CD8, IgT and antibody levels were assessed at 1, 3, and 7-day(s) post challenge (dpc). The results revealed that antibody levels in the group (ONB + CS) + Vac were consistently higher than the other groups post immersion immunization and oral booster, along with elevated expression of immune-related genes after challenge with V. harveyi. Ultimately, this group demonstrated a significantly higher relative percent survival (RPS) of 63 % ± 10.5 %, showcasing the potential of the ONB-CS-Vac complex as a promising immersion vaccination strategy for enhancing antigen uptake, stimulating immunological responses, and improving survival of Asian seabass against vibriosis.
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  • 文章类型: Journal Article
    弧菌与活海鲜有关,因为它们是土著海洋微生物区系的一部分。在亚洲,由弧菌引起的食源性感染。很常见。近年来,副溶血性弧菌已成为所有报道的食物中毒暴发的主要原因。因此,研究了卤代酸及其33种衍生物对副溶血弧菌的抗菌作用。化合物3,5-二碘-2-甲氧基苯基硼酸(DIMPBA)和2-氟-5-碘苯基硼酸(FIPBA)具有抗菌和抗生物膜活性。DIMPBA和FIPBA对浮游细胞生长的最小抑制浓度为100μg/mL,并以剂量依赖性方式防止生物膜形成。两种碘硼酸都可以减少影响运动性的几种毒力因子,菌毛凝集,疏水性,和吲哚合成。因此,这两种活性卤代酸阻碍了浮游和生物膜细胞的增殖。此外,这些化合物具有有效抑制鱿鱼和虾模型表面生物膜形成的潜力。
    Vibrios are associated with live seafood because they are part of the indigenous marine microflora. In Asia, foodborne infections caused by Vibrio spp. are common. In recent years, V. parahaemolyticus has become the leading cause of all reported food poisoning outbreaks. Therefore, the halogenated acid and its 33 derivatives were investigated for their antibacterial efficacy against V. parahaemolyticus. The compounds 3,5-diiodo-2-methoxyphenylboronic acid (DIMPBA) and 2-fluoro-5-iodophenylboronic acid (FIPBA) exhibited antibacterial and antibiofilm activity. DIMPBA and FIPBA had minimum inhibitory concentrations of 100 μg/mL for the planktonic cell growth and prevented biofilm formation in a dose-dependent manner. Both iodo-boric acids could diminish the several virulence factors influencing the motility, agglutination of fimbria, hydrophobicity, and indole synthesis. Consequently, these two active halogenated acids hampered the proliferation of the planktonic and biofilm cells. Moreover, these compounds have the potential to effectively inhibit the presence of biofilm formation on the surface of both squid and shrimp models.
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